Eupatilin

Verticillium wilt, a significant pathogen affecting cotton, has historically been challenging to control, posing a substantial threat to the sustainable development of the cotton industry. This study demonstrates that resistance to Verticillium dahliae in cotton can be enhanced by treating the roots with an ethyl acetate extract (EAAA) extracted from Artemisia argyi. The mechanisms by which EAAA activates immunity in cotton were elucidated by examining the expression levels of resistance genes post-treatment, evaluating salicylic acid (SA) and jasmonic acid (JA) levels, analyzing transcriptome data, and employing virus-induced gene silencing (VIGS) technology. Additionally, pot experiments were conducted to validate the efficacy of EAAA in controlling Verticillium wilt. The flavonoid content in EAAA was qualitatively analyzed using Ultra-Performance Liquid Chromatography coupled with Tandem Mass Spectrometry (UPLC-MS/MS), identifying three specific flavonoids that were further screened to verify their roles in activating cotton immunity. Cotton plants treated with EAAA exhibited reduced leaf chlorosis and browning in the vascular bundles. Genes involved in SA and JA synthesis and signaling in the root system were highly expressed, resulting in increased levels of SA and JA. Transcriptome analysis revealed that most upregulated differentially expressed genes were primarily enriched in the Mitogen-Activated Protein Kinase (MAPK) signaling pathway. Two specific genes, RLK and MAPKKK18, were identified through VIGS technology as key regulators of the immune pathway in cotton. The flavonoid monomer activation experiment demonstrated that eupatilin, hispidulin, jaceosidin, and a mixture of these three could induce the expression of cotton-related resistance genes. Collectively, these findings provide a research basis for the development of EAAA as a natural plant immune-inducing agent against cotton Verticillium wilt.

Gratiola officinalis L. (hedge hyssop), a medicinal plant of the Scrophulariaceae family, has diuretic, purgative, and vermifuge properties. It is used as a herbal tea to treat chronic gastroenteritis, renal colic, jaundice, and intestinal worms. Previously, we have found that an extract from G. officinalis is nontoxic and has antitumor, antioxidant, antimicrobial, antiinflammatory, anticachexic, and other properties. Our aims in this study were to separate the G. officinalis extract into individual fractions, to identify the most biologically active fractions, and to examine the chemical composition of these fractions and their biological activity toward A498 renal carcinoma cells.

The G. officinalis extract was fractionated by reversed-phase high-performance liquid chromatography, and each fraction was tested for antitumor activity. The active fractions were characterized by UV–visible electron spectral analysis, circular dichroism analysis, Fourier transform infrared spectroscopy, high-performance liquid chromatography, electrospray ionization tandem mass spectrometry, and nuclear magnetic resonance spectroscopy.

Two antitumor-active fractions of a flavonoid nature were isolated and chromatographically purified. On the basis of the nuclear magnetic resonance data, the aglycone fragment of the main component of one fraction was found to be structured as 2-(3,4-dimethoxyphenyl)-7-hydroxychroman-4-one, or 3',4'-dimethoxy-7- hydroxyflavanone.

The antitumor effect of the most active fraction containing 7-O-glucoside of apigenin, glycoside 7,3'-di-O-luteolin and trace amounts of eupatilin against renal carcinoma A498 cells was manifested in its cytotoxic, cytostatic, apoptotic and autophagosomal activities. In addition, we found 3-(1-2)-glucoside of soyaspogenol B, which is a pentacyclic triterpenoid in the structure.